| 2001 |
CARD11 (CARMA1) is a CARD-containing MAGUK family scaffold protein whose CARD domain directly binds the CARD domain of BCL10; overexpression of CARD11 induces BCL10 phosphorylation and NF-κB activation in cells. |
Co-immunoprecipitation, domain mapping, overexpression assays in cell lines |
The Journal of biological chemistry |
High |
11278692 11356195
|
| 2002 |
CARD11 is required for TCR-induced NF-κB activation but not for NFAT or AP-1 activation; CARD11 acts upstream of the IKK complex and cooperates with BCL10 in a CARD domain-dependent manner. |
Somatic mutagenesis to generate CARD11-deficient T cell line, RNAi rescue experiments, reconstitution assays |
Nature immunology / The EMBO journal |
High |
12154356 12356734
|
| 2002 |
CARMA1 is constitutively associated with lipid rafts; TCR stimulation induces physical association of CARMA1 with the TCR and BCL10, and BCL10 translocates into lipid rafts upon TCR engagement. A CARMA1 mutant defective in BCL10 binding acts as dominant negative for TCR-induced NF-κB activation. |
Co-immunoprecipitation, lipid raft fractionation, dominant-negative overexpression, IL-2 production assay |
Nature immunology |
High |
12154360
|
| 2003 |
Genetic inactivation of CARD11 in mice results in complete block in T and B cell immunity with selective defect in JNK and NF-κB activation; CARD11 is required for antigen receptor- and PKC-mediated proliferation and cytokine production, as well as TLR4-mediated B cell proliferation. |
Genetic knockout mice, lymphocyte proliferation assays, cytokine production assays, signaling assays |
Immunity |
High |
12818157 12818158 12867038
|
| 2003 |
CARD11 knockout mice show that CARD11 requires its CARD domain to engage downstream signaling; mice expressing a CARDless CARMA1 (ΔCard) have defective NF-κB activation in B and T lymphocytes and impaired B cell development with absent CD5+ peritoneal B cells. |
Knock-in mice expressing CARDless CARMA1, NF-κB activation assays, B cell development analysis |
Current biology |
High |
12867037
|
| 2003 |
CARMA1 is required for Bcl10 recruitment to clustered TCR complexes and lipid rafts; CARMA1-deficient T cells have defective NF-κB activation and impaired IL-2 production, and CARMA1 is required for BCR and CD40-mediated B cell proliferation. |
CARMA1 knockout mice, immunofluorescence, lipid raft fractionation, reconstitution |
Current biology |
High |
12867038
|
| 2004 |
CARMA1 is constitutively associated with the cytoplasmic membrane; this membrane association is essential for CARMA1 function. Upon CD3/CD28 costimulation, CARMA1 recruits PKCθ, BCL10, and IKKβ into lipid rafts of the immunological synapse, and a CARMA1 mutant defective in membrane association (L808P) fails to rescue NF-κB activation. |
Subcellular fractionation, dominant-negative and reconstitution assays in CARMA1-deficient JPM50.6 T cells, lipid raft isolation, co-immunoprecipitation |
Molecular and cellular biology |
High |
14673152
|
| 2004 |
CARMA1 controls entry of IKK into lipid raft aggregates and the central region of the immune synapse downstream of PKCθ; CARMA1 is dispensable for overall immunological synapse formation and TCR redistribution but essential for IKK activation. |
CARMA1 knockout mice, peptide-specific T cell activation system, immunofluorescence at immunological synapse, IKK kinase assay |
The Journal of experimental medicine |
High |
15520247
|
| 2004 |
MALT1 is recruited to lipid rafts of the immunological synapse in a CARMA1-dependent manner; MALT1 associates independently with both BCL10 and CARMA1, forming a trimolecular CBM complex. A MALT1 deletion mutant blocks CD3/CD28-induced NF-κB activation. |
Co-immunoprecipitation in CARMA1-deficient T cells, lipid raft fractionation, dominant-negative overexpression, NF-κB reporter assay |
The Journal of biological chemistry |
High |
14754896
|
| 2004 |
CARMA1 physically associates with IKKγ/NEMO and participates in an inducible complex containing IKKγ/NEMO, BCL10, and IKKα/β kinases. Expression of the NEMO-binding region of CARMA3 exerts dominant negative effect on BCL10-mediated NF-κB activation. |
Co-immunoprecipitation, dominant-negative overexpression, NF-κB reporter assay |
The Journal of biological chemistry |
Medium |
15184390
|
| 2005 |
PKCβ phosphorylates CARMA1, and BCR engagement leads to progressive recruitment of CARMA1 into lipid rafts and association with PKCβ. PKCβ and PKCθ phosphorylate identical serine residues (S564, S649, S657) in the CARMA1 linker region; mutation of S564 and S657 ablates CARMA1 functional activity, whereas linker deletion causes constitutive, PKC-independent NF-κB activation. |
In vitro kinase assay, site-directed mutagenesis, lipid raft fractionation, co-immunoprecipitation, NF-κB reporter assay |
Immunity |
High |
16356855 16356856
|
| 2005 |
PKCθ phosphorylates CARMA1 in vitro and in vivo on Ser552 in the linker region; this phosphorylation is required for recruitment of downstream signaling components to the immunological synapse and for TCR-induced NF-κB activation. |
In vitro phosphorylation assay, site-directed mutagenesis (S552A), reconstitution in CARMA1-deficient T cells, immunological synapse imaging |
Immunity |
High |
16356856
|
| 2005 |
PKCβ mediates BCR-induced IKK activation by phosphorylating CARMA1, which facilitates interaction between CARMA1 and TAK1; IKK is recruited to the CARMA1-BCL10-MALT1 complex in a PKCβ-dependent manner. |
Co-immunoprecipitation, PKCβ-deficient DT40 B cells, in vitro reconstitution, IKK kinase assay |
The Journal of experimental medicine |
High |
16301747
|
| 2006 |
CaMKII is redistributed to the immune synapse following T cell activation; CaMKII phosphorylates CARMA1 on Ser109, and this phosphorylation facilitates the interaction between CARMA1 and BCL10 and is critical for NF-κB activation. |
In vitro kinase assay, site-directed mutagenesis, co-immunoprecipitation, immunofluorescence at immune synapse, NF-κB reporter assay |
Molecular and cellular biology |
Medium |
16809782
|
| 2006 |
IKKβ is required for initial CBM complex formation; after engaging IKKβ/MALT1/BCL10 with CARMA1, IKKβ phosphorylates BCL10 at the C-terminus, which negatively regulates BCL10/MALT1 association and BCL10-mediated IKKγ ubiquitination. Mutation of IKKβ phosphorylation sites on BCL10 enhances NF-κB target gene expression. |
IKKβ-deficient T cells (epistasis), in vitro kinase assay, site-directed mutagenesis, co-immunoprecipitation, cytokine expression assay |
Molecular cell |
High |
16818229
|
| 2006 |
CARMA1 selectively regulates JNK2 but not JNK1 downstream of TCR signaling; BCL10 is inducibly associated with JNK2 and serves as a scaffold to assemble JNK2, MKK7, and TAK1, and this regulates c-Jun protein levels. |
CARMA1-deficient and BCL10-deficient T cells, co-immunoprecipitation, kinase assay, immunoblot for c-Jun |
Immunity |
High |
17189706
|
| 2007 |
PKCβ mediates phosphorylation of CARMA1 on Ser668, which is essential for BCR-mediated CARMA1-BCL10-MALT1 complex assembly and IKK activation. IKKβ also contributes to CARMA1-BCL10-MALT1 complex formation through a feedback phosphorylation of CARMA1. |
Site-directed mutagenesis, phospho-specific antibodies, co-immunoprecipitation, PKCβ-deficient B cells, IKK kinase assay |
The Journal of experimental medicine |
High |
18086859
|
| 2007 |
The cytoplasmic tail of CD26 interacts with CARMA1 in T cells; caveolin-1 ligation of CD26 recruits a complex of CD26, CARMA1, BCL10, and IKK to lipid rafts, leading to NF-κB activation. |
Co-immunoprecipitation, lipid raft fractionation, NF-κB activation assay |
The Journal of biological chemistry |
Medium |
17287217
|
| 2007 |
CARMA1 coiled-coil domain mediates constitutive oligomerization of CARMA1; disruption of CC1 sub-domain abolishes CARMA1 localization while disruption of CC2 inhibits CARMA1 self-association. Both impair TCR-induced NF-κB activation. |
Co-immunoprecipitation, site-directed mutagenesis, subcellular localization imaging, NF-κB reporter assay |
The Journal of biological chemistry |
Medium |
17428801
|
| 2007 |
ADAP directly associates with CARMA1, and ADAP deficiency impairs TCR-stimulated CARMA1-BCL10-MALT1 complex assembly; a specific region of ADAP is required for CARMA1 association and NF-κB activation but not ADAP-dependent adhesion regulation. |
ADAP-deficient mouse T cells, co-immunoprecipitation, NF-κB nuclear translocation assay, domain mapping |
Science |
Medium |
17478723
|
| 2008 |
The region between the coiled-coil and PDZ domains of CARD11 functions as an inhibitory domain (ID) that controls CARD11 association with multiple signaling cofactors including BCL10, TRAF6, TAK1, IKKγ, and caspase-8 through interactions requiring both CARD and coiled-coil domains. TRAF6 and caspase-8 associate with CARD11 in a signal-inducible manner dependent on ID neutralization. |
RNA interference rescue assay, co-immunoprecipitation in BCL10- and MALT1-deficient cells, site-directed mutagenesis, NF-κB reporter assay |
Molecular and cellular biology |
High |
18625728
|
| 2008 |
Oncogenic CARD11 coiled-coil domain mutations (found in 9.6% of ABC DLBCL) cause constitutive NF-κB activation and enhance NF-κB activity upon antigen receptor stimulation when introduced experimentally into lymphoma cell lines. |
Tumor sequencing, experimental overexpression of mutant CARD11 in lymphoma cell lines, NF-κB activation assay |
Science |
High |
18323416
|
| 2009 |
Cbl-b E3 ligase promotes monoubiquitination of CARMA1, which disrupts its complex formation with BCL10 without affecting CARMA1 protein stability; this mechanism contributes to NKT cell anergy induction. |
Cbl-b knockout mice, in vitro ubiquitination assay, co-immunoprecipitation, IFN-γ production and tumor rejection assays |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
19815501
|
| 2009 |
CARMA1 is required for development of thymic FoxP3+ regulatory T cells in a T cell-intrinsic manner; the CBM complex controls an early checkpoint enabling generation of thymic Treg precursors. Constitutive STAT5 activity or BCL2 overexpression cannot rescue Treg development in CARMA1-deficient mice. |
CARMA1 knockout mice, adoptive transfer, transgenic rescue experiments, flow cytometric analysis of Treg development |
Journal of immunology / PLoS biology |
High |
19260764 19454668
|
| 2009 |
PP2A regulatory subunit Aα (PPP2R1A) interacts with CARMA1 and removes PKCθ-dependent phosphorylation at Ser645 (Ser649 in mouse) of CARMA1; maintaining Ser645 phosphorylation by PP2A knockdown augments CBM complex formation, NF-κB activation, and cytokine production in T cells. |
Co-immunoprecipitation, in vitro dephosphorylation assay, siRNA knockdown, phospho-specific antibodies, CBM complex immunoprecipitation |
The EMBO journal |
High |
21157432
|
| 2009 |
Serine 649 phosphorylation in the CARMA1 linker down-regulates CARMA1 activity; unlike early activating phosphorylations (S657, S564), S649 phosphorylation occurs later and is maintained for longer post-stimulation, and depends partly on novel PKC activity. CARMA1 S649A mutation leads to increased NF-κB and JNK activation at low stimulation levels. |
Site-directed mutagenesis, phospho-specific antibody generation, reconstitution in CARMA1-deficient DT40 B cells, PKCβ-deficient B cells, NF-κB and JNK activation assays |
Journal of immunology |
Medium |
19917688
|
| 2009 |
CARMA1 undergoes K48-linked polyubiquitination and proteasome-dependent degradation upon antigen receptor activation; the SH3 and GUK domains contain the main ubiquitin acceptor sites, and deletion of the Hook domain between SH3 and GUK induces constitutive ubiquitination. Elimination of MAGUK ubiquitination sites elevates basal and inducible NF-κB and JNK activation. |
Site-directed mutagenesis, ubiquitination assay, proteasome inhibitor treatment, NF-κB and JNK activation assays |
Molecular and cellular biology |
Medium |
20008554
|
| 2009 |
HPK1 kinase interacts with CARMA1 in a TCR stimulation-dependent manner and phosphorylates the CARMA1 linker region at S549, S551, and S552; CARMA1 S551A and S549A/S551A mutants fail to restore HPK1-mediated and TCR-mediated NF-κB activation in CARMA1-deficient T cells. |
In vitro kinase assay, site-directed mutagenesis, co-immunoprecipitation, reconstitution in CARMA1-deficient T cells, IL-2 expression assay |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
19706536
|
| 2010 |
Oncogenic CARD11 mutations F123I and L225LI induce CARD11 hyperactivity by disrupting autoinhibition by the inhibitory domain (ID); these mutations disrupt ID-mediated intramolecular interactions, bypass the requirement for ID phosphorylation during TCR signaling, and selectively enhance CARD11 affinity for BCL10. |
Site-directed mutagenesis, co-immunoprecipitation, NF-κB reporter assay, intramolecular interaction assay |
Biochemistry |
High |
20799731
|
| 2010 |
Calcineurin (calcium-dependent phosphatase) positively regulates TCR-induced NF-κB by controlling CBM complex formation; calcineurin interacts with the CBM complex, dephosphorylates BCL10 in vitro and in vivo, and its inhibition (cyclosporin A, FK506) or knockdown impairs CBM complex assembly and NF-κB activation. |
Calcineurin siRNA knockdown, in vitro dephosphorylation assay, co-immunoprecipitation, NF-κB reporter assay, calcium chelation |
The Journal of biological chemistry |
Medium |
21199863
|
| 2011 |
OX40 (TNFR family member) assembles a signalosome containing TRAF2, RIP, IKKα/β/γ, CARMA1, MALT1, BCL10, and PKC in membrane microdomains independent of TCR engagement; CARMA1 and PKC recruitment is required for strong NF-κB activation downstream of OX40. |
Co-immunoprecipitation, lipid raft fractionation, dominant-negative and siRNA approaches, NF-κB activation assay |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
21282629
|
| 2012 |
CARD11 gain-of-function mutations (coiled-coil domain) in BENTA patients cause spontaneous CARMA1 aggregation and constitutive NF-κB activation in B cells, while rendering T cells less responsive to antigen receptor-induced activation. |
Whole transcriptome sequencing, transfection of mutant constructs, NF-κB activation assay, patient lymphocyte analysis |
The Journal of experimental medicine |
High |
23129749
|
| 2012 |
A CARD11 LATCH domain functions with the CARD to promote CARD11 autoinhibition; gain-of-function mutations in the LATCH or CARD disrupt inhibitory domain binding, promote BCL10 association, and induce BCL10 ubiquitination, NF-κB activation, and human lymphoma cell survival. |
High-throughput quantitative signaling screen, site-directed mutagenesis, co-immunoprecipitation, NF-κB reporter assay, lymphoma cell survival assay |
Molecular and cellular biology |
Medium |
23149938
|
| 2013 |
The reconstituted CBM signalosome forms a helical filamentous assembly in which substoichiometric CARMA1 nucleates BCL10 filaments through CARD-CARD interactions; BCL10 filament formation is highly cooperative and threshold is sensitized by oligomerized CARMA1. Structure-guided mutagenesis confirmed interfaces for BCL10 filament assembly and MALT1 activation in vitro and NF-κB activation in cells. |
In vitro reconstitution, cryo-electron microscopy, X-ray crystallography, NMR, structure-guided mutagenesis, NF-κB activation assay |
Molecular cell |
High |
24074955
|
| 2013 |
Carma1 (with MALT1 but not BCL10) is required for optimal TCR-dependent mTOR/mTORC1 activation in T cells; MALT1 catalytic activity is required for proliferation and increased metabolic flux of activated CD4+ T cells. |
Carma1-deficient, MALT1-deficient, BCL10-deficient T cells; mTORC1 phosphorylation assays, metabolic flux assays, proliferation assays |
Science signaling |
Medium |
24917592
|
| 2013 |
STUB1 (E3 ubiquitin ligase) constitutively interacts with CARMA1, and upon TCR stimulation promotes K27-linked ubiquitination of CARMA1; this ubiquitination is required for CARMA1-mediated NF-κB activation and IL-2 production. |
Co-immunoprecipitation, RNAi knockdown, ubiquitin linkage-specific assay, NF-κB reporter assay, overexpression |
European journal of immunology |
Medium |
23322406
|
| 2013 |
USP9X deubiquitinase interacts with BCL10 of the CBM complex and removes TCR-induced ubiquitin chains from BCL10, which facilitates CARMA1 association with BCL10-MALT1; USP9X knockdown reduces NF-κB activation and T cell function. |
Co-immunoprecipitation, in vitro deubiquitination assay, USP9X knockdown chimeric mice, adoptive transfer |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
23690623
|
| 2015 |
SH3 and GUK domain interactions within CARMA1 (both intra- and inter-molecular) are required for activation-induced assembly of CARMA1 microclusters at the immunological synapse; disruption of these interactions abolishes CARMA1 clustering, NF-κB activation, and constitutive clustering in ABC-DLBCL cells with oncogenic CARMA1. |
Site-directed mutagenesis, live cell imaging at immunological synapse, co-immunoprecipitation, NF-κB reporter assay in ABC-DLBCL lines |
Nature communications |
High |
25602919
|
| 2016 |
CARD11 inhibitory domain (ID) contains four repressive elements (REs) that function cooperatively with redundancy to prevent spontaneous NF-κB activation; each RE contributes to closed inactive CARD11 state and prevention of BCL10 binding. Oncogenic mutations in CARD, LATCH, or coiled coil can perturb intramolecular interactions mediated by multiple REs simultaneously. |
Site-directed mutagenesis of multiple RE elements, intramolecular interaction assay, co-immunoprecipitation, NF-κB reporter assay |
The Journal of biological chemistry |
High |
26884334 26884335
|
| 2016 |
TCR signaling induces linear (Met1-linked) polyubiquitination of BCL10 (Lin(Ub)n-BCL10), which is required for BCL10 association with NEMO/IKKγ; CARD11 promotes Lin(Ub)n-BCL10 generation by co-recruiting BCL10 with HOIP (LUBAC subunit) via the coiled-coil domain. This requires BCL10 lysines 17, 31, and 63, MALT1, and CARD11 autoinhibitory elements. |
Ubiquitin linkage-specific assays, co-immunoprecipitation, site-directed mutagenesis, MALT1/BCL10-deficient cells, NF-κB reporter assay |
The Journal of biological chemistry |
High |
27777308
|
| 2017 |
Heterozygous loss-of-function CARD11 mutations exhibit dominant-interfering activity on antigen receptor-induced NF-κB and mTORC1 activation; mTORC1 and IFN-γ production defects are partially rescued by glutamine supplementation, indicating CARD11 is required for glutamine import into T cells. |
Transfection of mutant constructs in T cell lines, patient T cell functional assays, NF-κB and mTORC1 signaling assays, metabolic rescue with glutamine |
Nature genetics |
High |
28628108
|
| 2018 |
Cryo-EM structure of BCL10 CARD filament at 4.0-Å resolution redefines CARD-CARD interaction interfaces; BCL10 polymerizes unidirectionally with CARMA1 serving as nucleation hub for star-shaped filamentous networks. MALT1 cooperatively interacts with BCL10 filaments and immediately dimerizes in the BCL10 scaffold; TRAF6 cooperatively decorates CBM filaments forming higher-order assemblies for downstream signaling. |
Cryo-EM, time-lapse confocal microscopy, in vitro polymerization assay, electron microscopy of MALT1/TRAF6 interactions |
Proceedings of the National Academy of Sciences of the United States of America |
High |
29382759
|
| 2019 |
The CARD11 autoinhibited state is maintained by an extensive interface between its CARD and coiled-coil domains; disruption of this interface causes hyperactivation in cells and formation of BCL10-templating filaments in vitro. Oncogenic CARD11 mutations break CARD-coiled-coil autoinhibitory contacts to relieve autoinhibition. |
X-ray crystallography (CARD9 as structural proxy for CARD11 mechanism), in vitro filament formation assay, cell-based NF-κB hyperactivation assay, structure-guided mutagenesis |
Nature communications |
High |
31296852
|
| 2013 |
Homozygous loss-of-function CARD11 mutations in humans cause profound combined immunodeficiency with abrogated canonical NF-κB pathway activation in lymphocytes after antigen receptor or PMA stimulation; truncated CARD11 protein is defective in antigen receptor signaling and NF-κB activation in reconstitution assays. |
Whole-exome sequencing, reconstitution assays, flow cytometry, NF-κB activation assays in patient lymphocytes |
The Journal of allergy and clinical immunology / The Journal of allergy and clinical immunology |
High |
23374270 23561803
|
| 2016 |
Oncogenic CARMA1 recruits β-catenin and its destruction complex (APC, AXIN1, CK1α, GSK3β) via a proteomic interaction; this recruitment is independent of CBM complex formation or constitutive NF-κB activation and promotes β-catenin stabilization, augmenting WNT/TCF-LEF transcriptional responses and IL-10 expression. |
Proteomic approach in BJAB cells expressing oncogenic CARMA1, co-immunoprecipitation, β-catenin stabilization assay, TCF/LEF reporter assay |
Oncogene |
Medium |
26776161
|
| 2010 |
GAKIN (kinesin-3 family member) interacts with CARD11 in a signal-dependent manner, competes with BCL10 for CARD11 association, and negatively regulates TCR signaling to NF-κB; GAKIN dynamically localizes to the immunological synapse and regulates redistribution of CARD11 from the central to a distal synapse region. |
Expression cloning screen, co-immunoprecipitation, live cell imaging at immunological synapse, NF-κB reporter assay |
Molecular cell |
Medium |
21145487
|
| 2015 |
RNF181 E3 ubiquitin ligase interacts with CARD11, reduces BCL10 levels via ubiquitin ligase activity, and inhibits antigen receptor signaling to NF-κB downstream of CARD11; RNF181 limits proliferation of human DLBCL cells dependent on CARD11 signaling. |
Bioluminescence resonance energy transfer-based interaction cloning (BRIC), co-immunoprecipitation, in vitro E3 ligase assay with BCL10 as substrate, NF-κB reporter assay, lymphoma cell proliferation assay |
Molecular and cellular biology |
Medium |
26711259
|
| 2023 |
CARD11 gain-of-function mutations directly increase transcription of the anti-apoptotic gene BCL2A1, leading to resistance to venetoclax; MALT1 protease inhibition reduces BCL2A1 expression and induces synergistic cell death with BCL2 inhibition regardless of CARD11 mutational status. |
Single-cell RNA-sequencing, integrative genomic analysis, functional cell line studies, in vitro and in vivo MALT1 inhibitor combination assays |
Blood |
Medium |
37562004
|